Fig. 4

Immunoelectrophoretic analysis of C3 in NHS incubated with fractions S2 and S5. a Positive control (C+) with zymosan (31.25 μL, 1 mg/mL) and NHS (100 μL, 1:2), and negative control (C-) with PBS (50 μL) and NHS (100 μL, 1:2). b Fraction S2 (50 μL, A₂₈₀ ~ 0.2) with SHN (100 μL, 1:2). c Fraction S5 (50 μL, A₂₈₀ ~ 0.16) with NHS (100 μL, 1:2). All mixtures were incubated for 60 min in a water bath at 37 °C. The plates were dried at room temperature, stained with 0.5 % Ponceau and bleached with 10 % acetic acid. Electrophoretic conditions: first dimension – four hours, 140 V at 15 mA and 10 W; second dimension – 14 h at 15 mA and 10 W. d Figures were manipulated to remove the background highlighting the presence of one peak for C- and two peaks for C+, S2 and S5 assays